title: A high-throughput screen for tuberculosis progression.
abstract: One-third of the world population is infected with Mycobacterium tuberculosis and multi-drug resistant strains are rapidly evolving. The noticeable absence of a whole organism high-throughput screening system for studying the progression of tuberculosis is fast becoming the bottleneck in tuberculosis research. We successfully developed such a system using the zebrafish Mycobacterium marinum infection model, which is a well-characterized model for tuberculosis progression with biomedical significance, mimicking hallmarks of human tuberculosis pathology. Importantly, we demonstrate the suitability of our system to directly study M. tuberculosis, showing for the first time that the human pathogen can propagate in this vertebrate model, resulting in similar early disease symptoms to those observed upon M. marinum infection. Our system is capable of screening for disease progression via robotic yolk injection of early embryos and visual flow screening of late-stage larvae. We also show that this system can reliably recapitulate the standard caudal vein injection method with a throughput level of 2,000 embryos per hour. We additionally demonstrate the possibility of studying signal transduction leading to disease progression using reverse genetics at high-throughput levels. Importantly, we use reference compounds to validate our system in the testing of molecules that prevent tuberculosis progression, making it highly suited for investigating novel anti-tuberculosis compounds in vivo.
authors.affiliation: ZF-screens B.V., Leiden, The Netherlands. h.p.spaink@biology.leidenuniv.nl
authors.forename: Ralph
authors.initials: R
authors.lastname: Carvalho
authors.forename: Jan
authors.initials: J
authors.lastname: de Sonneville
authors.forename: Oliver W
authors.initials: OW
authors.lastname: Stockhammer
authors.forename: Nigel D L
authors.initials: ND
authors.lastname: Savage
authors.forename: Wouter J
authors.initials: WJ
authors.lastname: Veneman
authors.forename: Tom H M
authors.initials: TH
authors.lastname: Ottenhoff
authors.forename: Ron P
authors.initials: RP
authors.lastname: Dirks
authors.forename: Annemarie H
authors.initials: AH
authors.lastname: Meijer
authors.forename: Herman P
authors.initials: HP
authors.lastname: Spaink
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000302:isolation & purification / Q000627:therapeutic use; D015415:Biomarkers / Q000032:analysis*; D004195:Disease Models, Animal; D018450:Disease Progression; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004625:Embryo, Nonmammalian; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009165:Mycobacterium Infections, Nontuberculous / Q000175:diagnosis / Q000188:drug therapy / Q000473:pathology; D019910:Mycobacterium marinum / Q000502:physiology; D011379:Prognosis; D014376:Tuberculosis / Q000175:diagnosis* / Q000188:drug therapy / Q000473:pathology; D015027:Zebrafish / Q000196:embryology / Q000254:growth & development / Q000502:physiology
chemical_list: D000995:Antitubercular Agents; D015415:Biomarkers